Plastic litter colonization in a brackish water environment.

Transitional waters, including coastal ponds, represent unique environments. These distinct ecosystems are often among the most severely polluted systems due to intensive human activities. Our study marks the first evaluation of aquatic invertebrates associated with plastic litter in two brackish ponds. We collected 43 items of plastic litter (including bottles and disposable plastics) during the winter and spring of 2022. Most of plastic litter (76.8 %) was colonized by aquatic invertebrates. A total of 495 individuals were observed on the plastic litter, with the number of individuals ranging from 1 to 54 (average = 13.4). The most abundant taxa were from the families Gammaridae, Serpulidae, and Sphaeromatidae. Invertebrates colonized both the external and internal surfaces of the plastic bottles. Plastic bottles trapped 25 % of the total biota. The internal entrapped taxa were mainly represented by gammarids, molluscs egg mass, and sphaeromatids. Open bottles could potentially serve as temporary or permanent traps for invertebrates colonizing their internal surfaces. We argue that the dispersal of species driven by plastics is possible but limited, just as water exchanges with the seas are limited. These brackish lakes could be transit areas, but more importantly, they could be hotspots for plastic litter.

Monitoring of anthropogenic microplastic pollution in antarctic fish (emerald rockcod) from the Terranova Bay after a quarter of century.

Monitoring the occurrence of microplastic contamination in the Antarctic area is the key to implement policy measures for waste regulations in the research stations. Antarctic fish Trematomus bernachii is a suitable species for establishing microplastic contamination and for investigating changes over time in the concentration and type of microplastics in the Antarctic region. In this paper a total of 78 fish, caught during the 37th Italian Antarctic expedition (2021-2022) in the Ross Sea (Antarctica) were analysed. Different microfibers and dyes were identified by Raman spectroscopy and the results were compared with those obtained for fish sampled in 1998. Differences in polymer type emerged with a predominance of synthetic fibers with respect to natural ones. These changes appear to be related to the increased human activities in the Antarctica over the last twenty years and highlights the need to improve the environmental sustainability of the numerous research stations operating throughout that area.

Polymer Blends Based on 1-Hexadecyl-3-methyl Imidazolium 1,3-Dimethyl 5-Sulfoisophthalate Ionic Liquid: Thermo-Mechanical, Surface Morphology and Antibacterial Properties.

In this study, antibacterial polymer blends based on Polyvinyl Chloride (PVC) and Polystyrene-Ethylene-Butylene-Styrene (SEBS), loaded with the ionic liquid (IL) 1-hexadecyl-3-methyl imidazolium 1,3-dimethyl 5-sulfoisophthalate (HdmimDMSIP) at three different concentrations (1%, 5%, and 10%), were produced. The IL/blends were characterized by their thermo-mechanical properties, surface morphology, and wettability. IL release from the blends was also evaluated. The agar diffusion method was used to test the antibacterial activity of the blends against Staphylococcus epidermidis and Escherichia coli. Results from thermal analyses showed compatibility between the IL and the PVC matrix, while phase separation in the SEBS/IL blends was observed. These results were confirmed using PY-GC MS data. SEM analyses highlighted abundant IL deposition on PVC blend film surfaces containing the IL at 5-10% concentrations, whereas the SEBS blend film surfaces showed irregular structures similar to islands of different sizes. Data on water contact angle proved that the loading of the IL into both polymer matrices induced higher wettability of the blends' surfaces, mostly in the SEBS films. The mechanical analyses evidenced a lowering of Young's Modulus, Tensile Stress, and Strain at Break in the SEBS blends, according to IL concentration. The PVC/IL blends showed a similar trend, but with an increase in the Strain at Break as IL concentration in the blends increased. Both PVC/IL and SEBS/IL blends displayed the best performance against Staphylococcus epidermidis, being active at low concentration (1%), whereas the antimicrobial activity against Escherichia coli was lower than that of S. epidermidis. Release data highlighted an IL dose-dependent release. These results are promising for a versatile use of these antimicrobial polymers in a variety of fields.

Anthropogenic microparticles in the emerald rockcod Trematomus bernacchii (Nototheniidae) from the Antarctic.

Anthropogenic microparticles (AMs) were found for the first time in specimens of Trematomus bernacchii collected in 1998 in the Ross Sea (Antarctica) and stored in the Antarctic Environmental Specimen Bank. Most of the identified AMs were fibers of natural and synthetic origin. The natural AMs were cellulosic, the synthetic ones were polyester, polypropylene, polypropylene/polyester, and cellulose acetate. The presence of dyes in the natural AMs indicates their anthropogenic origin. Five industrial dyes were identified by Raman spectroscopy with Indigo occurring in most of them (55%). Our research not only adds further data to the ongoing knowledge of pollution levels in the Antarctic ecosystem, it provides an interesting snapshot of the past, highlighting that microplastics and anthropogenic fiber pollution had already entered the Antarctic marine food web at the end of the '90 s. These findings therefore establish the foundations for understand the changes in marine litter pollution over time.

Shallow-Water Hydrothermal Vents as Natural Accelerators of Bacterial Antibiotic Resistance in Marine Coastal Areas.

Environmental contamination by heavy metals (HMs) poses several indirect risks to human health, including the co-spreading of genetic traits conferring resistance to both HMs and antibiotics among micro-organisms. Microbial antibiotic resistance (AR) acquisition is enhanced at sites anthropogenically polluted by HMs, as well as in remote systems naturally enriched in HMs, such as hydrothermal vents in the deep sea. However, to date, the possible role of hydrothermal vents at shallower water depths as hot spots of microbial AR gain and spreading has not been tested, despite the higher potential risks associated with the closer vicinity of such ecosystems to coasts and human activities. In this work, we collected waters and sediments at the Panarea shallow-water hydrothermal vents, testing the presence of culturable marine bacteria and their sensitivity to antibiotics and HMs. All of the bacterial isolates showed resistance to at least one antibiotic and one HM and, most notably, 80% of them displayed multi-AR on average to 12 (min 8, max 15) different antibiotics, as well as multi-HM tolerance. We show that our isolates displayed high similarity (≥99%) to common marine bacteria, affiliating with Actinobacteria, Gammaproteobacteria, Alphaproteobacteria and Firmicutes, and all displayed wide growth ranges for temperature and salinity during in vitro physiological tests. Notably, the analysis of the genomes available in public databases for their closest relatives highlighted the lack of genes for AR, posing new questions on the origin of multi-AR acquisition in this peculiar HM-rich environment. Overall, our results point out that shallow-water hydrothermal vents may contribute to enhance AR acquisition and spreading among common marine bacteria in coastal areas, highlighting this as a focus for future research.

Microplastics in the bogue, Boops boops: A snapshot of the past from the southern Tyrrhenian Sea.

The present investigation focuses on Boops boops specimens gathered in the Gulf of Patti in 2010. Providing a snapshot from the past, this paper represents, chronologically, the first record of microplastic ingestion in the Mediterranean bogue. The plastic abundance and composition in gastrointestinal tracts of the bogue was assess, in order to improve the knowledge on spatial-temporal variability of microplastics pollution in the Mediterranean basin and in particular, in the southern Tyrrhenian Sea. In a total of 65 specimens, 180 particles of plastic (2.8 items/specimens), mainly belonging to microplastics class, were found. Fragments (63%) and fibres (30%) were the predominant shape categories. Eleven polymers were identified: polypropylene and polyethylene were the most abundant. Several synthetic polymers belonging to the class of elastomers were also observed. The study area is strongly influenced by the absence of trawl fishing activities and a low mixing level of the seabed that, together with the confluence of different watercourses and the presence of different kind of anthropic impact, including motorway, could make it a 'waste disposal site'. Finally, our results suggest the usefulness to retrieve older samples to better understand spatial-temporal changes in marine litter pollution over time.

Characterization of Five Psychrotolerant Alcanivorax spp. Strains Isolated from Antarctica.

Five psychrotolerant Alcanivorax spp. strains were isolated from Antarctic coastal waters. Strains were screened for molecular and physiological properties and analyzed regarding their growth capacity. Partial 16S rDNA, alk-B1, and P450 gene sequencing was performed. Biolog EcoPlates and the API 20E test were used to evaluate metabolic and biochemical profiles. Bacterial growth in sodium acetate was determined at 4, 15, 20, and 25 °C to evaluate the optimal temperature. Furthermore, the ability of each strain to grow in a hydrocarbon mixture at 4 and 25 °C was assayed. Biosurfactant production tests (drop-collapse and oil spreading) and emulsification activity tests (E24) were also performed. Concerning results of partial gene sequencing (16S rDNA, alk-B1, and P450), a high similarity of the isolates with the same genes isolated from other Alcanivorax spp. strains was observed. The metabolic profiles obtained by Biolog assays showed no significant differences in the isolates compared to the Alcanivorax borkumensis wild type. The results of biodegradative tests showed their capability to grow at different temperatures. All strains showed biosurfactant production and emulsification activity. Our findings underline the importance to proceed in the isolation and characterization of Antarctic hydrocarbon-degrading bacterial strains since their biotechnological and environmental applications could be useful even for pollution remediation in polar areas.

Is Wild Marine Biota Affected by Microplastics?

The present review provides detailed information on the adverse effects of MPs on wild marine organisms, including tissue damage, fish condition, oxidative stress, immune toxicity, and genotoxicity. A bibliometric analysis was carried out on CiteSpace (version 6.1.R3) (Drexel University, Philadelphia, PA, USA) to verify how many papers studied the effects on wild marine species. The results showed a total of 395 articles, but only 22 really presented data on the effects or impacts on marine biota, and of these, only 12 articles highlighted negative effects. This review shows that the observed effects in wild organisms were less severe and milder than those found in the experimental conditions. The knowledge of negative effects caused by direct ingestion of microplastics in wild animals is still limited; more efforts are necessary to fully understand the role of MPs and the adverse effects on wild marine organisms, the ecosystem, and human health.

Thermo-mechanical, antimicrobial and biocompatible properties of PVC blends based on imidazolium ionic liquids.

The aim of this study was the development of antimicrobial polyvinylchloride (PVC) blends loaded with 0.1-10% (w/w) of the ILs 1-hexadecyl-3-methylimidazolium 1,3-dimethyl 5-sulfoisophthalate (HdmimDMSIP) and 1-octyloximethyl-3-methylimidazolium hexafluorophosphate (OOMmimPF6). The synthetized ILs were characterized by 1HNMR, MALDI-TOF, DSC and TGA. PVC/ILs films were obtained by solvent casting.Thermal and mechanical properties (tensile stress TS and elongation at break EB), morphology by SEM, surface wettability, antimicrobial activity, cytotoxicity and ILs release in sterile water from PVC/ILs film blends were determined. Results demonstrated that the presence of both ILs in PVC formulation slightly affected thermal and mechanical properties of blends. The loading of both ILs into PVC matrix made PVC/ILs films hydrophilic, especially at the highest concentration of HdmimDMSIP. The PVC/ILs blends displayed antibacterial activity up to ILs lowest concentrations (0.1-0.5%). The inhibition of Escherichia coli growth was lower than that showed toward Staphylococcus epidermidis. The addition of 10% ILs concentration resulted excessive as demonstrated by accumulation of ILs on film surfaces (SEM) and ILs high release from PVC/ILs blends during the first day of water immersion. Biocompatibility studies highlighted that the addition of low amounts of both ILs into PVC matrix is not cytotoxic for mouse fibroblast cells (L929), supporting their potential use for biomedical porposes.

Plastics occurrence in juveniles of Engraulis encrasicolus and Sardina pilchardus in the Southern Tyrrhenian Sea.

We report the presence of microplastics on the external surface and in the gastrointestinal tract of white late-larval and juvenile stages (fry) of clupeid fishes caught in the Southern Tyrrhenian Sea. The average highest number of plastics debris was recorded on Sardina pilchardus (0.53 items/specimen); a lower average number of items was observed for Engraulis encrasicolus (0.26 items/specimen). The plastics were characterized by fibers that differed in shape, colour and composition. Polyester, polypropylene, polyacrylonitrile, polyethylene, polyamide, nylon, rayon and polyurethane segments were detected by Raman and FTIR spectroscopies. Traces of organic components and dyes, compounds that are generally included in the polymer matrix to modify its base properties, were also identified on microplastics. Our results raise concerns for the potential transfer of synthetic materials through the marine food web and into humans, given the prominent role of S. pilchardus and E. encrasicolus within the food web as main food source for many marine species.

The antioxidant power of horseradish, Armoracia rusticana, underlies antimicrobial and antiradical effects, exerted in vitro.

Armoracia rusticana (AR) was tested for antimicrobial and antioxidants power. The compound demonstrated to inhibit fish pathogens such as Vibrio anguillarum, V. harvey, V. alginolyticus, Aeromonas hydrophila, A. salmonicida, Photobacterium damselae subspecie piscicida, Tenacibaculum marinum and Pseudomonas anguilliseptica,. The total phenolic content and the reducing power resulted higher in the water extract of AR, respect to the hydroalcoolic. In vitro test demonstrated that AR significantly protect cells against death, induced by oxidative stress.

Quali-quantitative analysis of plastics and synthetic microfibers found in demersal species from Southern Tyrrhenian Sea (Central Mediterranean).

This study highlights plastics occurrence in five demersal fish species from the Southern Tyrrhenian Sea: the Red mullet Mullus barbatus barbatus, the Piper gurnard Trigla lyra, the Blackmouth catshark Galeus melastomus, the Lesser spotted dogfish Scyliorhinus canicula and the Brown ray Raja miraletus. Overall, 125 fish were examined: 21 Red mullets, 16 Piper gurnards, 75 Blackmouth catsharks, 72 Dogfish and 1 Brown ray. The percentage of fish with ingested plastics was 14.4% with 0.24 items per specimen. The majority of the debris were fibers and the application of infrared and Raman spectroscopy allowed the identification and discrimination of plastic and non-plastic fibers. The plastic debris isolated were mainly microplastics (94.1%), while macroplastics occurrence was very low (5.9%). The plastics were identified as polypropylene, Teflon, nylon, kraton G (triblock copolymer) and polyethylene. Also cellulose was detected. S. canicula was the species with the highest number of plastic pollutants.

Plastics occurrence in the gastrointestinal tract of Zeus faber and Lepidopus caudatus from the Tyrrhenian Sea.

The present study investigates the occurrence of plastic pollution in two commercially important marine teleosts (Zeus faber and Lepidopus caudatus) from the northern coasts of Sicily (Tyrrhenian Sea). Plastics occurrence in the gastrointestinal tract was higher in Lepidopus caudatus (78.1%) than Zeus faber (51.4%). Debris characterization, carried out by micro-Raman spectroscopy, allowed identified the main types of found polymers as: polypropylene (PP), polyamide (PA), nylon and, to a lesser extent, polyethylene (PE). Of the two fish species studied, the silver scabbardfish appeared to be the more vulnerable to plastic ingestion. Our study represents a starting point that may pave the way for future investigation of the fate, accumulation and transfer of plastic debris to upper trophic levels, to verify their potential toxicity and to better understand strategies to mitigate this phenomenon.

Detection of artificial cellulose microfibers in Boops boops from the northern coasts of Sicily (Central Mediterranean).

Pollution deriving from textile wastes, including industrial and household waste, is recently of great interest due to their environmental impacts. Anthropogenic and synthetic fibers are responsible for negative effects on the quality of water and soil, and, also, their presence damages plant and animal health. In this work, the authors revealed the occurrence of man-made cellulose fibers in specimens of Boops boops from the Northern Sicilian coasts. Bogue was chosen as target species as it has been used as an indicator within the European Marine Strategy Framework Directive (MSFD 2008/56/EC) in order to value the "microplastics status" in the stomach contents. Of the 30 specimens examined, 63.3% of these had ingested fibers items. The number of fibers ranged from 1 to 10 per specimens with an average of 2.7 items/specimen. Fibers length ranged from 0.5 to 30 mm, most of them were black (95%), and a small percentage was red (5%). The ingestion of man-made cellulose fibers, observed for the first time, in Boops boops in the Mediterranean Sea wake-up call and it should attract the attention of the EU for new guidelines where this new type of contaminant is classified harmful as well as plastics.

First record of microplastics ingestion by European hake MERLUCCIUS MERLUCCIUS from the Tyrrhenian Sicilian coast (Central Mediterranean Sea).

A sample of 67 European hake Merluccius merluccius were examined to highlight the ingestion of microplastics in the Tyrrhenian Sea. In all samples, 31 black fibres were found in the stomach contents corresponding to 46.3% of the specimens. The data presented here could be important for the implementation of the EU Marine Strategy Framework Directive in Mediterranean waters.

Isolation of a novel gene from Photobacterium damselae subsp. piscicida and analysis of the recombinant antigen as promising vaccine candidate.

Photobacterium damselae subsp. piscicida (PDP) is the causative agent of fish pasteurellosis, a bacterial disease causing important losses in marine aquaculture. Vaccines against the pathogen can be a way to control the infection and avoid antibiotic treatments. However, a satisfactory protective vaccine against fish pasteurellosis is not commercially available. In this study, a biotechnogical approach based on reverse vaccinology has been used to identify potential vaccine candidates for the development of a recombinant subunit vaccine. Genome sequencing of clones from a genomic cosmid library of PDP and in silico selection of the surface exposed proteins were the initial steps in vaccine candidate identification. From 370 open reading frames (ORF) eight potential antigens were selected, expressed as recombinant proteins and purified. These vaccine candidates were used to generate specific polyclonal antibodies in mice. Each antibody was then screened in vitro by inhibition adherence assay of live PDP on chinook salmon embryo cells (CHSE-214). A lipoprotein, found to be involved in the adherence of the bacterium to epithelial cells and annotated as PDP_0080, was then selected. The recombinant protein was further investigated in fish vaccination and challenge experiments to assess its ability to protect sea bass, Dicentrarchus labrax, against PDP infection. Immunisation with PDP_0080 recombinant protein elicited high specific antibody titres. Furthermore, the survival rate of fish immunized with the 25 µg dose of protein was significantly higher compared to the control group. The results of the study suggest that the PDP_0080 protein could be a promising candidate for the design of a recombinant vaccine against pasteurellosis.

Comparison of 16SrDNA and toxR genes as targets for detection of Vibrio anguillarum in Dicentrarchus labrax kidney and liver.

Vibrio anguillarum is a pathogen that causes high mortality in marine and freshwater fish. The aim of this study was to develop a real-time PCR assay for identification and quantification of V. anguillarum in fish tissue. The assay was carried out using two target genes, 16SrDNA and toxR, to evaluate the influence of differences in the operon copy number in quantitative assessment, both in pure cultures of V. anguillarum serovar O1 (strain 975/I), as a reference, and in the liver and kidney of a sea bass (Dicentrarchus labrax) specimen. Real-time PCR analysis showed high specificity for both target genes, with a detection limit of approximately 1-10 bacterial cells per reaction in pure culture and 10/100 V. anguillarum cells per reaction in fish tissue, which corresponds to 2 × 10(2)/2 × 10(3) cells g(-1) fish tissue. Moreover, both genes showed high specificity but differing sensitivity due to the different operon copy number; as a result, it is possible to target the high copy number gene to improve sensitivity. Our results suggest that the protocol we tested can be used as a sensitive and specific molecular method for the detection of the fish pathogen V. anguillarum in fish tissue.

First report on antibody response of Seriola dumerilii (Risso 1810) challenged with Listonella anguillarum.

The greater amberjack, Seriola dumerilii (Risso 1810) is a semi-pelagic fish and a worldwide species; it is considered a promising candidate for the diversification of Mediterranean aquaculture. In this paper an experimental injection with Listonella (Vibrio) anguillarum was performed to study the immune response of S. dumerilii. Antibody titres to L. anguillarum O1 were determined with indirect-ELISA at different times over a period of 42 days. Results showed that the antibody levels against L. anguillarum were significantly higher in the challenged fish compared to the control. They started developing since the 5th day reaching the highest peak on day 20 after injection, indicating a fast response of the immune system. The observed antibody titre was very high versus L. anguillarum if compared to other fish species.

Evaluation of different DNA-based fingerprinting methods for typing Photobacterium damselae ssp. piscicida.

This study evaluates the effectiveness of three different molecular techniques, repetitive extragenic palindromic PCR (REP-PCR), enterobacterial repetitive intergenic consensus sequence PCR (ERIC-PCR) and the random amplified polymorphic DNA (RAPD-PCR) for rapid typing of Photobacterium damselae ssp. piscicida strains isolated from different species of marine fish and geographic areas. The results obtained by the three methods showed that RAPD and ERIC-PCR were more discriminative for suitable rapid typing of Ph. damselae ssp. piscicida than REP-PCR. The analysis of DNA banding patterns generated by both molecular methods (RAPD and ERIC-PCR) clearly separated the strains into two main groups that strongly correlated with their geographic origin. Moreover, the REP-PCR analysis was less reproducible than the RAPD and ERIC-PCR methods and does not allow the establishment of genetic groups. RAPD and ERIC-PCR constitute valuable tools for molecular typing of Ph. damselae ssp. piscicida strains, which can be used in epidemiological studies of photobacteriosis infections.

Evaluation of different DNA-based fingerprinting methods for typing Photobacterium damselae ssp. piscicida

This study evaluates the effectiveness of three different molecular techniques, repetitive extragenic palindromic PCR (REP-PCR), enterobacterial repetitive intergenic consensus sequence PCR (ERIC-PCR) and the random amplified polymorphic DNA (RAPD-PCR) for rapid typing of Photobacterium damselae ssp. piscicida strains isolated from different species of marine fish and geographic areas. The results obtained by the three methods showed that RAPD and ERIC-PCR were more discriminative for suitable rapid typing of Ph. damselae ssp. piscicida than REP-PCR. The analysis of DNA banding patterns generated by both molecular methods (RAPD and ERIC-PCR) clearly separated the strains into two main groups that strongly correlated with their geographic origin. Moreover, the REP-PCR analysis was less reproducible than the RAPD and ERIC-PCR methods and does not allow the establishment of genetic groups. RAPD and ERIC-PCR constitute valuable tools for molecular typing of Ph. damselae ssp. piscicida strains, which can be used in epidemiological studies of photobacteriosis infections.